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PCR

PCR

Maxam-Gilbert Sequencing

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

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Updated: Jun 9, 2026

Identification of Functional Protein Regions Through Chimeric Protein Construction

Identification of Functional Protein Regions Through Chimeric Protein Construction

Published on: January 8, 2019

Chimera construction using multiple-template-based sequential PCRs.

Qiang Shan¹, Joseph W Lynch

¹Queensland Brain Institute, University of Queensland, Brisbane, QLD, Australia. q.shan@uq.edu.au

Journal of Neuroscience Methods

|September 7, 2010

Summary

This summary is machine-generated.

We developed a simplified protein chimera construction protocol using a multiple-template polymerase chain reaction (PCR) approach. This method significantly improves success rates and reduces complexity compared to traditional techniques.

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CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

Published on: June 25, 2017

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Last Updated: Jun 9, 2026

Identification of Functional Protein Regions Through Chimeric Protein Construction

Identification of Functional Protein Regions Through Chimeric Protein Construction

Published on: January 8, 2019

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

Published on: June 25, 2017

Area of Science:

Molecular Biology
Protein Engineering

Background:

Protein chimeras are valuable tools for studying protein structure-function relationships.
Traditional chimera construction methods are often complex and have low success rates.

Purpose of the Study:

To describe a novel, simplified protocol for constructing protein chimeras.
To overcome the limitations of traditional chimera construction techniques.

Main Methods:

Introduced a "multiple-template" concept for PCR-based chimera construction.
Eliminated the need for restriction sites and purification of intermediate PCR products.
Utilized two to three simple PCRs followed by general subcloning.

Main Results:

Achieved a nearly 100% success rate in chimera construction.
Significantly reduced the procedural complexity compared to traditional methods.
The protocol does not require specific restriction sites.

Conclusions:

The described protocol offers a highly efficient and simplified method for protein chimera construction.
This technique is broadly applicable for protein engineering and functional studies.
The high success rate makes this method suitable for various research applications.